Abstract
Genetically encoded FRET based biosensors allow one to visualize the spatial and temporal evolution of specific enzyme activities in live cells. We have previously reported the creation of a FRET based biosensor specific for Zeta-Associated Protein -70 kD (ZAP-70) (Randriamampita et al., 2008), a Syk family protein tyrosine kinase. ZAP-70 is essential for early T cell receptor (TCR) signaling events, T lymphocyte development and has also been implicated in integrin mediated T lymphocyte migration. In order to facilitate the study of ZAP-70 kinase activity during dynamic phenomena such as immunological synapse formation or cell migration, we have designed and prepared a second generation of ZAP-70 specific biosensors. Here we describe a novel biosensor named ROZA-XL, that displays a 3-4 times greater dynamic range than its predecessor and possesses a robust baseline FRET value when expressed in the Jurkat human T cell line. We demonstrate that the robust behavior of this biosensor allows for rapid analysis of TCR mediated of ZAP-70 kinase activity at a single cell level, as shown in a simple end point assay in which ROZA-XL expressing cells are allowed to interact with stimulatory anti-CD3epsilon coated coverslips.
References
Dec 26, 2001·Proceedings of the National Academy of Sciences of the United States of America·A Y TingR Y Tsien
Oct 3, 2002·The Journal of Cell Biology·Stephen C BunnellLawrence E Samelson
Dec 4, 2002·Nature Reviews. Molecular Cell Biology·Jin ZhangRoger Y Tsien
Apr 7, 2004·Biochemistry·Jon C D HoutmanLawrence E Samelson
Feb 8, 2005·Nature Biotechnology·Annalee W Nguyen, Patrick S Daugherty
Sep 19, 2008·Proceedings of the National Academy of Sciences of the United States of America·Mingxing OuyangYingxiao Wang
Jan 16, 2009·Immunity·Claire ConcheClotilde Randriamampita
Mar 18, 2009·Immunological Reviews·Byron B Au-YeungArthur Weiss
Aug 12, 2009·Molecular and Cellular Biology·R G BakerG A Koretzky
May 11, 2010·Cold Spring Harbor Perspectives in Biology·Haopeng WangArthur Weiss
Nov 3, 2010·Nature Immunology·Byron B Au-YeungArthur Weiss
Jan 5, 2011·Blood·Rachel EvansNancy Hogg
Mar 6, 2012·Journal of Cell Science·Alexander A SmoligovetsJay T Groves
Nov 15, 2012·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Rebekah R Bartelt, Jon C D Houtman
Jul 25, 2013·Science Signaling·Rafael D FritzOlivier Pertz
Oct 30, 2013·Biotechnology Journal·Clotilde Randriamampita, Annemarie C Lellouch
Mar 7, 2014·ELife·Misty R JenkinsGillian M Griffiths
Aug 8, 2014·Biophysical Journal·Omer DushekVahid Shahrezaei
Citations
Oct 27, 2015·Sensors·Bernhard HochreiterJohannes A Schmid
Jul 8, 2016·Annals of Biomedical Engineering·Kaitao LiCheng Zhu
Jan 30, 2019·Frontiers in Immunology·Julia M LauferDaniel F Legler
Mar 20, 2018·Cell Structure and Function·Gembu MaryuKazuhiro Aoki
Jul 29, 2019·The Journal of Membrane Biology·Laurent Limozin, Pierre-Henri Puech
Oct 20, 2020·Wiley Interdisciplinary Reviews. Systems Biology and Medicine·Xinyuan ZhangKeyue Shen
Jun 7, 2021·Current Opinion in Cell Biology·Kendelle J MurphyPaul Timpson
Aug 21, 2021·Nature Communications·Longwei LiuYingxiao Wang